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Systematic Bacteriology Actinomycetes

Systematic Bacteriology Actinomycetes

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Published by Koji Elegado
Guide to Actinomycete identification
Guide to Actinomycete identification

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Published by: Koji Elegado on Feb 03, 2014
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INTERNATIONAL JOURNAL OF SYSTEMATIC BACTERIOLOGY Vol. 16,
No.
3 July 1966 pp. 313-340 METHODS FOR CHARACTERIZATION
OF
STREPTOMYCES
SPECIES'
E. B. Shirling and D. Gottlieb Department of Botany and Bacteriology Ohio Wesleyan University, Delaware, Ohio and Department of Plant Pathology University of Illinois, Urbana, Illinois ABSTRACT. The methods used by collaborators in the International Streptomyces Project
(ISP)
for. emendation of descriptions of type and neotype strains of the genus Streptomyces (Actinomycetales) are presented. An international cooperative effort, now in progress,
is
directed toward collection of type cultures
of
the Strepto- myces species for deposition with the Centraalbureau voor Schimmelcultures (CBS), Baarn. From this center the ref- erence cultures will be supplied to other culture collections so that they are available throughout the world. An essential adjunct to this activity
is
the redescription of each type culture in terms of currently acceptable cri- teria and methods. Theurgent need for an authentic refer- ence collection, accompanied by standardized characteriza- tions for each species, has been pointed out by spokesmen for the several meetings and conferences which culminated in this project. (See, for example, Gottlieb, 1959, 1961; Kiister, 1959; Krasil'nikov, I961
.)
More than
40
investi- gatorst representing
17
countries are participating in this research. Each culture
is
described independently by three of these cooperating specialists in different laboratories before it
is
deposited in the reference collection. This project
is
supported in part by
a
research grant from the National Science Foundation,
U.
S.
A. The Subcommittee on Actinomycetes of the Committee on Taxonomy, A.
S.
M. and the Subcommittee on,Taxonomy
of
Actinomycetes
of
the International Committee on Bacteriological Nomenclature are co-sponsoring advisors. Participants in the 1964-1965 studies are listed on p. 338.
 
314
INTERNATIONAL JOURNAL This manual contains the criteria and methods adopted for the project.
It
reflects the results of two extensive co- operative studies directed toward selection of stable prop- erties and reproducible procedures for characterization of streptomycetes. One study conducted under the direction of the Subcommittee on Actinomycetes of the Committee on Taxonomy, American Society for Microbiology was reported by the Chairman, Dr.
D.
Gottlieb
1961).
A
similar
pre- liminary study on an international basis was reported for the Subcommittee on Taxonomy of Actinomycetes of the In- ternational Committee on Bacteriological Nomenclature by the Secretary, Dr.
E.
Kiister (1961,1964). The descriptive criteria are essentially the same as those included in the recommendations of this international subcommittee for descriptions of Actinomycetales appearing in patent appli- cations (Gottlieb, 1963). The methods in mimeographed form have been used successfully for the description of type and neotype strains of more than
200
named species sub- mitted to
ISP
collaborators during 1964 and 1965, and are
now
in use for a continuation of the project. Only minor editorial changes have been made except that the test for nitrate reduction (including medium
8,
Bacto-nitrate broth) has been omitted. This characteristic proved unstable and has been dropped from the study. It is hoped that the characterizations used in this manual will be included in future descriptions of Streptomyces spe- cies
so
that direct comparison can be made with descrip- tions for type cultures in the reference collections. MATERIALS AND GENERAL METHODS CULTURE MEDIA Prepare Difco%
4
dehydrated culture media
as
instructed on labels of containers.
If
the dehydrated media are not used, use formulas and instructions in this manual
as
a
guide to preparation of the media. All culture media described in this manual have been pre- pared especially for the
I.S.
P.
by Difco Laboratories
as
pref ormulat ed dehydrated media. This important cont ribu- tion by Dif co Laboratories
is
gratefully acknowledged. When Difco dehydrated media are used, instructions on labels supersede instructions in the manual. Difco Laboratories, Detroit, Michigan,
U.
S.
A. 48201
 
SYSTEMATIC BACTERIOLOGY 315. Sterilize culture media in the autoclave
at
121 C. Steri- lize loosely packed'tubes or flasks containing
less
than
500
mlfor 15 minutes; herilize larger quantities for 20 minutes. (Do not autoclave carbon compounds to be used in the carbon utilization tests. Special instructions for sterilizing these compounds are given with medium
9.
Adjust pH of media with NaOH or HC1 before addition of agar and before
steri-
lization. Trace salts solution
(Use
as
directed in
media
3,
4,
5
and 7
if
prepared from formulas. Do not add to the corres- ponding Difco dehydrated media.) FeS04. 7Hz0
....
.
.......
0.1 g MnClz*4H20
....
0.1 g ZnS04.7Hz0
............
0.1 g Distilled water
...........
100.
0
ml
Medium 1: Tryptone-yeast extract broth (Pridham and Gottlieb,
1948
Bacto-Tryptone (Difco) 5.0
g
Bacto-Yeast Extract (Difco)
......
3.0 g Distilled water 1.0
liter
pH 7.0 to 7.2 before autoclaving Dispense 5
ml
of broth into
test
tubes with
a
diameter One tube will be needed for of 20
mm
or more. growth from lyophile pellet. each culture studied. meyer flask (or 25
ml
into 125
ml
flask). will be used for preparation of washed inoculum (p. 322). One flask will be needed for each culture studied.
Use
these test tubes for initiating Dispense
50
ml
of the broth into each
250
ml
Erlen- These flasks Medium
2:
Yeast extract-malt extract agar (Pridham
eJ
-
l.,
1956-57) Bacto-Yeast Extract (Difco)
......
4.0
g Bacto-Malt Extract (Difco)
......
0.0 g Bacto-Dextrose (Difco)
.......
4.0 g Distilled water
...........
Adjust to pH
7.,3,
hen add Bacto agar
.............
20.0 g Liquefy agar by steaming
at
100 C for 15-20 minutes. 1.
0
liter

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